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Supplementary Figures 1 through 7 and Supplementary Tables 1 and 2 from <i>TET2</i> Mutations Affect Non-CpG Island DNA Methylation at Enhancers and Transcription Factor–Binding Sites in Chronic Myelomonocytic Leukemia

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posted on 2023-03-30, 23:11 authored by Jumpei Yamazaki, Jaroslav Jelinek, Yue Lu, Matteo Cesaroni, Jozef Madzo, Frank Neumann, Rong He, Rodolphe Taby, Aparna Vasanthakumar, Trisha Macrae, Kelly R. Ostler, Hagop M. Kantarjian, Shoudan Liang, Marcos R. Estecio, Lucy A. Godley, Jean-Pierre J. Issa
<p>7 supplementary figures, and 2 supplementary tables. Supplementary Figure 1. Scatter plots for DNA methylation levels analyzed by DREAM. Supplementary Figure 2. Difference in DNA methylation for 7 repeat classes by bisulphite-pyrosequencing for TET2-MT, TET2-WT cases, and normal blood samples. Supplementary Figure 3. Unsupervised hierarchical analyses for DNA methylation levels for all sites analyzed by DREAM. Supplementary Figure 4. Supervised hierarchical analysis for DNA methylation levels for tet2-DMCs. Supplementary Figure 5. Representative genomic landscapes for tet2-DMCs more methylated in TET2-MT in NCGIs. Supplementary Figure 6. Bar plots for the enrichment of tet2-DMCs in NCGI for regulatory including promoter flanking regions, promoters, enhancers, and CTCF binding sites defined by Ensembl Regulatory Build data. Supplementary Figure 7. p300-related genes are deregulated in TET2-MT. Supplementary Table 1. Primer sequences Supplementary Table 2. Number of unique usable tags obtained from DREAM</p>

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ARTICLE ABSTRACT

TET2 enzymatically converts 5-methylcytosine to 5-hydroxymethylcytosine as well as other covalently modified cytosines and its mutations are common in myeloid leukemia. However, the exact mechanism and the extent to which TET2 mutations affect DNA methylation remain in question. Here, we report on DNA methylomes in TET2 wild-type (TET2-WT) and mutant (TET2-MT) cases of chronic myelomonocytic leukemia (CMML). We analyzed 85,134 CpG sites [28,114 sites in CpG islands (CGI) and 57,020 in non-CpG islands (NCGI)]. TET2 mutations do not explain genome-wide differences in DNA methylation in CMML, and we found few and inconsistent differences at CGIs between TET2-WT and TET2-MT cases. In contrast, we identified 409 (0.71%) TET2-specific differentially methylated CpGs (tet2-DMCs) in NCGIs, 86% of which were hypermethylated in TET2-MT cases, suggesting a strikingly different biology of the effects of TET2 mutations at CGIs and NCGIs. DNA methylation of tet2-DMCs at promoters and nonpromoters repressed gene expression. Tet2-DMCs showed significant enrichment at hematopoietic-specific enhancers marked by H3K4me1 and at binding sites for the transcription factor p300. Tet2-DMCs showed significantly lower 5-hydroxymethylcytosine in TET2-MT cases. We conclude that leukemia-associated TET2 mutations affect DNA methylation at NCGI regions containing hematopoietic-specific enhancers and transcription factor–binding sites. Cancer Res; 75(14); 2833–43. ©2015 AACR.

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